The present invention relates to a device for binding molecules, molecular groups, molecular parts and/or cells contained in a liquid to target structures, as well as to a method for binding molecules, molecular groups, molecular parts and/or cells contained in a liquid to target structures.
The only method known so far for binding molecules, molecular groups, molecular parts and/or cells contained in a liquid to target structures is the one according to “Stamper and Woodruff (1976) J. Exp. Med 144, 828-833”. This method comprises the fixing of target structures to a surface and subsequently rehydrating them. Then, the target structures are coated with 100 to 200 μl of the liquid containing the molecules, groups of molecules, parts of molecules and/or cells. Incubation (e.g.: 30 min, 7° C.) thereof on a shaker or rotating table is followed by a rinsing step which will remove any non-binding matter.
One major disadvantage of this method is the fact that it is performed manually. Moreover, motion devices are used for obtaining the binding equilibrium which only poorly simulate the conditions prevailing in certain natural systems. Another shortcoming is that the rinsing step may destroy the often weak adherence of ingredients of the liquid to the target structures. Yet another disadvantage of this method is that it is not capable of stopping metabolic processes.
It is therefore the object of the present invention to provide a device and a method for simulating the short-term interactions occurring between molecules, molecular groups, molecular parts and/or cells in a liquid, on the one hand, and the target structures, on the other, at the same time allowing most of the metabolic processes to be stopped at any time.
This object is accomplished by a device and a method having the features of the independent claims.
Advantageous embodiments are described in the subclaims.
A device of the invention for binding molecules, molecular groups, molecular parts and/or cells contained in a liquid to target structures comprises at least one object holder with at least one target structure firmly held therein which is in contact with at least one duct made of at least one material, said duct including, at its one aperture, a liquid inlet and, at its other aperture, a liquid outlet, as well as at least one object holder thermostat used for cooling the respective object holder.
This is based on the inventive idea that flowing a liquid containing molecules, molecular groups, molecular parts and/or cells through a duct which is in contact with a target structure will cause said molecules, molecular groups, molecular parts and/or cells to bind to said target structure. Any non-binding matter can be removed via the second duct aperture. At the same time, the temperature of the object holder will be controlled by the object holder thermostat. Owing to the flowing motion, passing the liquid containing molecules, molecular groups, molecular parts and/or cells over the target structure is a better way of simulating natural systems with short-term interactions than by means of a prior art shaker or a prior art rotating table. A particular advantage of the invention is that it allows an interruption of metabolic processes at any time by decreasing the temperature of the object holder by means of the object holder thermostat. In doing so, the temperature of the object holder will be 2-10° C., in particular 4° C.
In an advantageous embodiment of the invention, the duct is formed by the object holder and paraffin-like sheet material. This allows the duct to widen, if necessary, preventing excessive pressure build-up within said duct.
In yet another advantageous embodiment of the invention, the discharged liquid will be collected in a vessel, thus preventing a loss of liquid which has already passed the duct, which liquid may be difficult and/or expensive to prepare.
In another advantageous embodiment of the invention, the temperature of the vessel will be cooled by a vessel thermostat, thus preventing any metabolic processes from occurring within said vessel and allowing the liquid or its ingredients to be used again.
A method of the invention for binding molecules, molecular groups, molecular parts and/or cells contained in a liquid to target structures comprises the following steps: (a) passing liquid through a duct and over an object holder carrying a target structure firmly held therein, (b) removing any non-binding matter via a second duct aperture and collecting such matter in a vessel, (c) cooling said object holder by means of an object holder thermostat, and (d) fixing the molecules, molecular groups, molecular parts and/or cells adhering to the object holder.
In yet another advantageous embodiment of the invention, the device is used in an automated method for determining molecular classes, molecular groups, molecular parts in a solid or liquid object according to U.S. patent application Ser. No. 09/353,942, the entire specification of which is incorporated herein by this reference. This will allow identification and characterization of the binding partners. It will be possible at the same time to identify and characterize the molecular classes, molecular groups and/or molecular parts required for the binding process. Such application will be advantageous in numerous areas of chemistry, biology, biochemistry and especially medicine. This will not only open up new possible ways of diagnosing, but will also provide a new test system for therapeutic approaches on a molecular level.